Induction of fibrinolysis

ABSTRACT

METHOD FOR INDUCING BIBRINOLYSIS IN MAMMALS, AND PHARMACEUTICAL COMPOSITIONS USEFUL THEREFOR, BY THE ACTION OF SPECIFIED ALKYLENEBIS (1,2,3,4-TERRAHYDROISOQUINOLINE) COMPOUNDS OR PHARMACEUTICALLY ACCEPTABLE ACID ADDITION SALTS THEREOF.

United States Patent Office 3,560,620 Patented Feb. 2, 1971 3,560,620INDUCTION F FIBRINOLYSIS Joseph M. Schor, Locust Valley, and NathanWeiner, Rego Park, N.Y., assignors to Endo Laboratories Inc. No Drawing.Filed Aug. 9, 1966, Ser. No. 571,195

Int. Cl. A61k 27/00 US. Cl. 424-258 16 Claims ABSTRACT OF THE DISCLOSUREMethod for inducing fibrinolysis in mammals, and pharmaceuticalcompositions useful therefor, by the action of specified alkylenebis[l,2,3,4-tetrahydroisoquinoline] compounds or pharmaceuticallyacceptable acid addition salts thereof.

This invention relates to means for inducing fibrinolytic activity inmammals. More particularly, it is directed to effecting high, sustained,useful levels of fibrinolytic activity in mammals by the action ofalkylenebis [1,2,3,4-tetrohydroisoquinoline] compounds of the followingFormula A, or pharmaceutically acceptable acid addition salts thereof,and to compositions containing such substances Formula A wherein:

The term pharmaceutically acceptable addition salt includes such saltsas: the mineral acid salts, e.g., the hydrochloride, hydrobromide,sulfate and phosphate; and organic acid salts such as the succinate,benzoate, acetate, p-toluene-sulfonate and benzenesulfonate, and othersformed from acids conventionally used in the pharmaceutical art.

In the formation of a blood clot, for example, athrombus, fibrinogen, asoluble plasma protein, is converted to the insoluble protein fibrin. Asthe fibrin is deposited it entraps blood cells within its meshwork toform a coagulum. In the case of a thrombus, the coagulum usuallyinterferes with the flow of blood through the vessel.

The defense of the living organism against such occurrence is the plasmaprotein called plasminogen which, under certain conditions, can beactivated by an activator whereby the plasminogen is converted to theprotein, plasmin. Plasmin possesses the property of efficientlydigesting and destroying fibrin (fibrinolysis). The fibrinolysis resultsin dissolution of the clot; and in the case of a thrombus, restores thepotency of the vessel.

Under normal conditions, the organism has low levels of acitvator in theblood stream. It is believed that small amounts of plasminogen areconstantly undergoing conversion to plasmin by the action of theactivator. However, from a quantitative viewpoint the amount ofactivator normally present is insuflicient to produce enough plasmin tolyse the relatively large amount of fibrin present in a clot such as athrombus.

The compounds of foregoing Formula A have the property of stimulatingthe production and/or release of relatively large quantities ofactivator activity into the blood stream. In essence, those compoundsinduce increased levels of activator which, in turn, produce high levelsof plasmin within the clot and this results in dissolution thereof.

Consequently, the compounds of Formula A can be used to dissolve fibrinwherever it is deposited in the organism, independent of the cause ofsuch deposition.

Fibrinolytic activity in vitro is manifested by many compounds such asthe aromatic sulfonic acids, derivatives of salicyclic acid, long chainfatty acids and halogenated unsaturated acids. The compounds of thepresent invention, more importantly, are active in vivo.

Fibrinolytic activity in vivo can be induced by nicotinic acid,procaine, phenylbutazone, acetylcholine, epinephrine, serotonin andhistamine. However, the effect of these compounds is of short-livedduration, i.e., of the order of minutes. The compounds of Formula A, incontrast thereto, can induce prolonged fibrinolytic activity, i.e., ofthe order of hours.

Some sulphonylureas and steroids can induce an increase in fibrinolyticactivity, but a lag period of the order of hours precedes the slowincrease in lyti cactivity. Compounds of this type cannot be used when asubstance is employed to effect thrombolytic therapy, because in suchinstance the activity must be rapidly induced to be effectice indissolving clots. The compounds of Formula A, in contrast thereto, arehighly effective for such use because they induce maximal lytic activitywithin minutes after administration.

Streptokinase, a streptococcal protein, has been used for thrombolysis,but the side effects of pyrogenicity and anaphylactic reactions havelimited its use. In contrast thereto, compounds of Formula A are free ofthese side reactions.

Urokinase, a protein isolated from human urine, has also been used forthrombolysis but the difficulties involved in accumulating largesupplies of the starting material, human urine, and the great cost ofpreparing the substance have proscribed its general and practicalutility.

Bacterial pyrogens have also been used to effect thrombolysis, but theseverity and unpredictability of the pyrogenic reactions have negatedtheir usefulness. The compounds of Formula A are not pyrogenic.

In short, the fibrinolytic compounds of Formula A are the firstsynthetic substances which demonstrate activity in mammals in vivo, arehighly potent, long-lasting, rapid in onset, readily prepared and sufferfrom none of the difiiculties associated with materials of naturalorigin.

The compounds of Formula A may be used to effect fibrinolysis of theclot in acute thrombosis. They may also be employed prophylactically tomaintain increased fibrinolytic activity or a long term basis and thusdiminish the icidence of new thrombotic episodes.

The alkylenebis [1,2,3,4-tetrahydroisoquinoline] compounds and theiracid addition salts present in the compositions of the invention, whenused in accordance with this invention, possess a high degree ofactivity and consequently only small amounts are required to producefibrinolytic effects in mammals. At these dose levels, side effects aregenerally negligible.

When substituents R and R in Formula A include hydroxy moieties,moderate to marked blood pressure lowering effects accompany theproduction of good fibrinolytic action.

Oxygenated functions are generally beneficial for maximum fibrinolyticactivity and with one preferred aspect of this invention there isincluded in R and R one or more lower alkoxy functions such as methoxy,isopropoxy, butoxy or methylenedioxy. These functions are most effectivewhen situated at the 6 and 7 positions of the tetrahydroisoquinolinering systems.

The alkylene chain, designated as alk, may vary within wide limits withretention of fibrinolytic potency. Activity is maximized, however, whenthe tetrahydroisoquinoline rings are separated by chains of from threeto ten carbon atoms. Examples of such chains are 2-propyltrimethylene,tetramethylene, 1,8-diethyloctamethylene and decamethylene.

The groups R R R and R in preferred compounds, represent hydrogen oralkyl groups of not more than four carbon atoms. Most preferred arethose compounds in which R R R and R are hydrogen or methyl.

The compounds of Formula A are all either known or, in view of theirstructures, are readily prepared by methods frequently described in theliterature. Generally speaking, their preparation comprises theinteraction of an appropriately substituted phenylethylamine with adicarboxyalkane, whereof the bridge between the carboxyl moietiescorresponds to the length of the alkylene chain desired. Thebis-phenylethylamide thus derived is then cyclized through dehydrationto an alkylene-bis-[3,4-dihydroisoquinoline]. The latter is thenconverted into a compound of Formula A through reduction to thecorresponding alkylenebis l,2,3,4-tetrahydroisoquinoline].

It will be apparent from a consideration of the synthetic processesinvolved in making these compounds that they are capable of occurring inisomeric forms. In the simplest instances, i.e., where R R R and R ofFormula A represent hydrogen, there are d1 and meso diastereoisomers.When additional asymmetric centers are introduced the situation becomesmore complex and more diastereoisomers are possible.

We have studied the fibrinolytic potencies of various diastereoisomersas well as the potencies of the diastereoisomeric mixtures from whichthey were derived. In each case the pure compounds and the mixtures werefound to be useful fibrinolytic agents. Formula A therefore embraces allof the isomers arising during the synthesis of these compounds. It isintended that the designation of the compounds of Formula A, by namesand by formulas throughout the specification and claims, shall be readto include all isomers and mixtures thereof unless otherwise specified.

An outstanding feature of the compounds of Formula A is their relativelack of toxicity. Although they are active at very low dose levels, itis possible to exceed minimum effective levels by wide margins withoutencountering serious adverse reactions. This permits the compounds to beused without concern about undesirable side effects, as are sofrequently encountered upon inadvertent overdosage of other substancesof great potency. For example, by intraperitoneal administration torats, compounds of Formula A were found to be effective in inducingfibrinolysis at 0.05 mg./kg. It was found, indeed, that the samecompounds could be administered at levels of 100 mg./kg. By intravenousadministration to rats, the compounds were effective at 0.05 ing/kg. andalso at 80 mg./kg. They were active by the oral route in rats at dosesof from 5 to 300 mg./kg. In dogs they were active by intraveneousadministration at 0.5 mg./kg. and also at 60 mg./kg. In humans they maybe administered parenterally in doses of from 0.1 mg./kg. to 50 ing/kg.These dose levels are illustrative and are not meant to limit theapplication of the invention. The veterinarian or physician may find itadvisable to adjust dosage in t light of individual recipient response.

The compositions of the invention contain analkylenebis-[1,2,3,4-tetrahydroisoquinoline] having Formula A or anon-toxic acid addition salt thereof together with a carrier. Thecarrier may be either a solid or liquid and the compositions can be inthe form of tablets, liquidfilled capsules, dry filled capsules,dragees, pills, aqueous solutions, non-aqueous solutions, jellies,suppositories, syrups, suspensions, sprays, powders and the like. Thecompositions can, and in many cases, do contain suitable preservatives,coloring and flavoring agents. Some examples of the carriers which canbe used in the preparation of the products of the invention are gelatincapsules, sugars such as lactose and sucrose; cellulose, methylcellulose and cellulose acetate phthalate; gelatin; talc; magnesiumstearate; vegetable oils such as peanut oil, cottonseed oil, sesame oil,olive oil, corn oil and oil of theobroma; liquid petrolatum,polyethylene glycol; glycerine; sorbitol; propylene glycol; ethanol;agar; water and isotonic saline.

In preparing the compositions of the invention for pharmaceutical uses,the conventional practices and precautions are used. The compositionsintended for parenteral administration must be sterile and this can beaccomplished either by using sterile ingredients and carrying out theproduction under asceptic conditions or by sterilizing the finalcomposition by one of the usual procedures such as milliporefilitration. Customary care should be exercised that no incompatiblecondition exists between the active component and the diulentpreservative or flavoring agent or in the conditions employed inpreparation of the compositions.

The compositions of the invention can be introduced into the mammal bythe oral, rectal, or parenteral route. This can be done by injecting theliquid preparations intraveneously, intramuscularly, intraperitoneally,or subcutaneously; by swallowing, in the cases of the solid and liquidpreparations, by local application to the mucous membrances, in the caseof jellies, suppositories, tablets and the like; by inhalation of spraysor mists of the liquid preparations and the like.

For a fuller understanding of the nature and objects of this invention,reference may be had to the following examples which are given merely asfurther illustrations of the invention and are not to be construed in alimiting sense. The temperatures are in degrees centigrade.

EXAMPLE 1 1, l '-tetramethylenebis[ l,2,3,4-tetrahydroisoquinoline]dihydrochloride A mixture of 35.2 g. of adipic acid di(2-phenylethyl)amide, M.P. 184l85, and 176 g. of polyphosphoric acid was heated at190193 for 1.25 hours. After cooling to the mixture was poured onto icealong with 20 ml. of concentrated hydrochloric acid. After washing withchloroform and benzene, the aqueous layer was made strongly alkalinewith aqueous sodium hydroxide, extracted with benzene, and the benzenesolution washed with water, dried, and purified by passage through analumina column. The residue of l,l-tetramethylenebis-[3,4-dihydroisoquinoline] remaining on evaporation of the benzene meltedat 100-102 after recrystallization from hexane-ethyl acetate.

To a solution of 44.2 g. of the 3,4-dihydro compound in 500 ml. ofethanol, 21.2 g. of sodium borohydride was added in portions withstirring. After the exothermic reaction had abated, the mixture wasrefluxed for 3 hours, and then evaporated to dryness in vacuo. Theresidue was taken up in water, extracted with chloroform, and theextract dried and evaporated to dryness. The resulting oil was dissolvedin absolute ethanol and treated with hydrogen chloride and ether. Thesolid product, representing an isomeric mixture of hydrochlorides,melted at 335- 338 (dec.). Fractional crystallization yielded one isomer(1.1) (A), M.P. 344-346 (decomposition) from water, and the secondisomeric product (B) (1.2), M.P. 266- 269 from ethanol.

EXAMPLE 2 1,1-hexamethylenebis 1,2,3,4-tetrahydroisoquinoline]dihydrochloride A mixture of 38 g. of suberic acid di-(Z-phenylethyl)amide, M.P. 165-166, and 300 g. of polyphosphoric acid was heated at 240for 1.5 hours. After cooling to 100- it was poured into 500 ml. of icewater containing ml. of concentrated hydrochloric acid. After washingwith benzene, the aqueous phase was made basic and extracted severaltimes with benzene. Concentration product 1,1-hexamethylenebis[3,4-dihydroisoquinoline], M.P. 9092 afterrecrystallization from acetonitrile.

A 30 g. sample of the bisdihydro compound was dissolved in ethanol and13 g. of sodium borohydride was added with stirring. After 10 minutes atroom temperature the mixture was heated under reflux for 1.5 hours. Theethanol was removed, water was added and the aqueous mixture wasextracted with chloroform. The chloroform solution was dried andevaporated. The oily residue was taken up in ethanol, treated withhydrogen chloride and cooled in ice. The resulting isomer mixture ofhydrochlorides melted at 275-285. Fractional crystallization yielded oneisomer (A) (2.1), M.P. 291-294 from methanolethyl acetate and the secondisomeric product (B) (2.2), M.P. 246-248 from ethanol.

EXAMPLE 3-1,1'-hexamethylenebis[7-chloro-1,2,3,4-tetrahydroisoquinoline]dihydrochloride By the procedude of Example 2, suberic acid di[2-(4-chloropheny1)ethyl] amide was converted into the mixture of isomeric(3.1) bis[7-chlorotetrahydroisoquinoline] dihydrochlorides.

EXAMPLE 4 1,1'-hexamethylenebis[7-chloro-1,2,3,4-tetrahydroisoquinoline]dihydrochloride A mixture of 8.24 g. of adipic aciddi[2-(3-methoxyphenyl)ethy1] amide, M.P. 152, ml. of phosphorusoxychloride, and 82 ml. of dry toluene was refluxed for 0.5 hour. Aftercooling in ice, the precipitate was filtered and washed with benzene andacetone. A solution of this solid in warm water was made stronglyalkaline with aqueous sodium hydroxide, and the product extracted intochloroform, washed with water and dried. Removal of the solvent gave1,1'-tetramethylenebis[3,4-dihydro- 6-methoxyisoquino1ine], M.P. 105106after recrystallization from cyclohexane. Reduction with sodiumborohydride as described in Example 1, followed by treatment withhydrogen chloride yielded the mixture of 11 and meso isomeric products,(4.1) M.P. 272-273".

EXAMPLE 5 1,1'-tetramethylenebis1,2,3,4-tetrahydro-6,7-methy1enedioxyisoquinoline] A mixture of 43 g. ofadipic acid di[2-(3,4-methylenedioxyphenyl)ethyl] amide, M.P. 207208,and 130 ml. of phosphorus oxychloride was heated under reflux for onehour. Excess phosphorus oxychloride was removed under reduced pressureand the residue was treated with water and then with alkali andchloroform. The chloroform was concentrated and the crystalline residuewas recrystallized from toluene to yield 1,l-tetramethylenebis[3,4-dihydro-6,7-methylenedioxyisoquinoline], M.P. 214- 215.

The dihydro compound (17 g.) was dissolved in hot dioxane (2 liters) andtreated dropwise with 6.4 g. of sodium borohydride in 200 ml. ofethanol. After the exothermic reaction subsided, the mixture was heatedunder reflux for an additional 3 hours and concentrated. The mixture ofdiasterioisomers (5.1) melted at 175-190.

6 EXAMPLE 6 1,1'-tetramethylenebis [7-benzyloxy-1,2,3,4-tetrahydro-6-methoxy-isoquinoline] dihydrobromide A mixture of 6.53 g. of adipic aciddi-[2-(4-benzyloxy-3-methoxyphenyl)ethyl] amide, M.P. 197, 20 ml. ofphosphorus oxychloride, and 65 m1. of dry toluene was refluxed for 1hour. After cooling the precipitate was washed with benzene and acetone.A solution of the precipitate in hot water was made strongly alkalinewith aqueous sodium hydroxide, and, after cooling in ice, theprecipitate was filtered and washed with water. Recrystallization fromabsolute ethanol gave 1,l-tetramethylenebis[7 benzyloxy 3,4dihydro-6-methoxyisoquinoline], M.P. 166. Reduction thereof with sodiumborohydride as described in Example 1, followed by treatment withhydrogen bromide yielded the mixture of (11 and meso isomerhydrobromides (6.1).

The following products are prepared in similar fashion:

(6.2) 1,1'-tetramethylenebis[7 allyloxy 1,2,3,4-tetrahydro-6-methoxyisoquinoline] (6.3) 1,1-tetramethylenebis[1,2,3,4tetrahydro 6- methoxy-7-phenoxyisoquinoline] EXAMPLE 71,1'-tetramethylenebis[1,2,3,4-tetrahydro-7-hydr0xy-6-methoxyisoquinoline] Reduction of1,1-tetramethylenebis[7-benzyloxy-3,4-dihydro-6-methoxyisoquinoline]described in Example 6, with 10% palladium on charcoal and hydrogen inacetic acid resulted in debenzylation to produce the mixture of isomeric7-hydroxy compounds, (7.1) M.P. 259-261".

EXAMPLE 8 1,1'-tetramethylenebis-[6-benzyloxy-7-butoxy-1,2,3,4-tetrahydroisoquinoline] dihydro chloride 2-(3-benzyloxy 4butoxyphenyl)-N-acetylethylamine was hydrolyzed with dilute sodiumhydroxide and the resulting amine was converted into the desired product(8.1) by the procedures of Example 6.

EXAMPLE 9 1,1'-(2-propyltrimethylene) 1,2,3,4-tetrahydroisoquinoline)1,2,3,4-tetrahydro-6,7-dimethoxyisoquinoline) 3-(3,4-dihydro 6,7dimethoxy-l-isoquinolylmethyl)- hexanoic acid and 2-phenylethylaminewere heated at 200 to produce the amide. Cyclization with phosphorusoxychloride followed by reduction with sodium borohydride as describedin Example 1 yielded the product.

EXAMPLE 1O1,1'-tetramethylenebis[1,2,3,4-tetrahydro-6,7,8-trimethoxyisoquinoline]dihydrochloride A mixture of 19.0 g. of adipic aciddi-[2-(3,4,5-trimethoxyphenyDethyl]amide, M.P. 174-175 57 ml. ofphosphorus oxychloride, and ml. of dry toluene was refluxed for 0.5 hourto give an oil which solidified on cooling. The solid was filtered,washed with benzene and acetone, and then dissolved in hot water andmade strongly 7 EXAMPLE 11 Ingredients: Mg. tablet 1,1'hexamethylenebis[l,2,3,4 tetraisoquinoline]dihydrochloride,isomerA 25Lactose USP (spray dried) 170 Starch USP Magnesium stearate USP 1Stearic acid USP 5 Flavor Q.s.

All the above ingredients were passed through 60 mesh sieve, blended for30 minutes and compressed directly into tablets on a suitable press at aweight of 211 mg. using a biconcave, scored punch.

'EXAMPLE 12 Ingredients: Mg./Capsule 1,1-hexamethylenebis[1,2,3,4-tetramethylene- 6,7-dihydroxyisoquinoline] dihydrobromideLactose USP 100 Magnesium stearate 1 Cab-O-Sil (amorphous silicondioxide) 5 These ingredients were combined, blended and passed through aNo. 1 screen of Fitzpatrick comminutor machine before encapsulating intoa two piece hard gelatin No. 3 capsule on a standard capsulating machineat a net weight of 126 mg.

EXAMPLE 13 Ingredients: Ampoule l,1-tetramethylenebis[1,2,3,4-tetrahydro-6,7- dimethoxyisoquinoline] dihydrochloride, isomer B mgPyrogen free water for injection ml 25 Mannitol N.F. To make thesolution isotonic.

The compound is added to the water and the solution made isotonic withmannitol. The resulting pH is 5.5. The

solution is filled into ampoules under sterile conditions and the sealedampoule is autoclaved. The ampoule contains 5 ml. of the 5 mg./ml.solution.

EXAMPLE 14 Ingredients: Ampoule l,1'-hexamethylenebis[1,2,3,4-tetrahydro-6,7

dimethoxy-3-methylisoquinoline] dihydrochloride mg 25 Pyrogen free waterfor injection ml 25 Dextrose N.F. To make the solution isotonic.

The compound is added to the water and the solution made isotonic withdextrose. The resulting pH is 5.5. The solution is filled into ampoulesunder sterile conditions and the sealed ampoule is autoclaved. Theampoule contains 5 ml. of the 5 mg./ml. solution.

EXAMPLE 15 Ingredients: Ampoule 1,1 -decamethylenebis 1,2,3,4-tetrahydro-6,7- dimethoxyisoquinoline] dihydrochloride dihydrate mg25 Pyrogen free water for injection ml 25 Fructose N.F. To make thesolution isotonic.

The compound is added to the water and the solution made isotonic withfructose. The resulting pH is 5 .5 The solution is filled into ampoulesunder sterile conditions and the sealed ampoule is autoclaved. Theampoule contains 5 ml. of the 5 mg./ml. solution.

EXAMPLE 16 Ingredients: Per ampoule1,1-tetramethylenebis[1,2,3,4-tetrahydro-6- methoxyisoquinoline] mg 25Peanut oil ml 5 Benzyl alcohol mg 5 The compound is dissolved in thepeanut oil containing the benzyl alcohol. The ampoules are filled understerile conditions. Each ampoule contains 5 ml. of 5 mg. of thecompound/ml.

EXAMPLE 17 Ingredients: Mg,/suppository1,l-hexamethylenebis[7-chloro-1,2,3,4-tetrahydroisoquinoline] 200 Cocoabutter c- Q.s.

The compound and cocoa butter are combined, mixed thoroughly and formedinto two gram suppositories.

EXAMPLE 18 Ingredients Mg./ suppository 1,1'-hexamethylenebis[1,2,3,4-tetrahydroisoquinoline], isomer B 200 Oil of theobroma Q.s.

The compound and oil of theobroma are combined, mixed thoroughly andformed into two gram suppositories.

EXAMPLE 19 Ingredients: Grams/liter 1,1-hexamethylenebis[1,2,3,4-tetrahydro-6,7-

dimethoxyisoquinoline] dihydrochloride 10 Granulated sugar 600 FlavorQ.s. Color Q.s. Sodium benzoate 1 'Deionized water Q.s.

All above ingredients are dissolved in water, combined and made up to avolume of one liter.

EXAMPLE 20 Ingredients: Mg./ tablet 1,1'-hexamethylenebis[1,2,3,4-tetrahydro-6,7

dimethoxyisoquinoline] dihydrochloride 50 Lactose USP (spray dried)Starch USP 20 Magnesium stearate USP 1 Stearic acid USP 5 Flavor Q.s.

These ingredients were passed through 60 mesh sieve, blended for 30minutes and compressed directly into tablets on a suitable tablet pressat a weight of 226 mg. using a biconcave, scored punch.

What is claimed is:

1. Method of treating a mammal to induce an increase in fibrinolyticactivity in a mammal in need of such treatment, which comprisesadministering to said mammal a fibrinolytically effective amount of acompound of the following Formula A or a pharmaceutically acceptableaddition salt thereof:

4. The method of claim 3 wherein said lower alkoxy is methoxy.

5. The method of claim 1 wherein said compound of Formula A is1,1alkylenebis[1,2,3,4-tetrahydro-hydroxyisoquinoline] 6. The method ofclaim '1 wherein said compound of Formula A is a1,1-alkylenebis[1,2,3,4-tetrahydro-lower alkoxy-hydroxy-isoquinoline] 7.The method of claim 1 wherein said compound of Formula A is a1,1'-alkylenebis[1,2,3,4-tetrahydro-methyl isoquinoline] 8. The methodof claim 1 wherein said compound of Formula A is1,1-alkylenebis[1,2,3,4-tetrahydro-methylenedioxy-isoquinolinc] 9.Method in accordance in claim 1 wherein the compound of Formula A is:1,1-tetramethylene-bis[1,2,31,4- tetrahydroisoquinoline] 10. Method inaccordance with claim 1 wherein the compound of Formulae A is:1,1-hexamethylene-bis[1,2, 3,4-tetrahydro-6-methoxyisoquinoline] 11.Method in accordance with claim 1 wherein the compound of Formula A is:1,1'-hexamethylene-bis[1,2, 3,4-tetrahydroisoquinoline] 12. Method inaccordance with claim 1 wherein the compound of Formula A is:1,1-tetramethylene-bis[1,2, 3,4-tetrahydro-6,7-dimethoxyisoquinoline]13. Method in accordance with claim -1 wherein the compound of Formula Ais: 1,1'-decamethylene-bis[1,2,3,4-tetrahydro-6,7-dimethoxyisoquinoline] References Cited UNITED STATESPATENTS 2,659,728 11/1953 Craig et al. 260-286 2,744,901 12/1953Nabenhauer 260-286 3,138,600 6/1964 Fancher et al. 260286 FOREIGNPATENTS 922,828 1/1'955 Germany 260-286 OTHER REFERENCES J. Chem. Soc.2010-21 (1929); Chemical Abstracts vol. 43 2624 (1949) (I); vol. 48 2254(1954) (II); Vol. 49 6957 (1955) (111); vol. 52 12870 (1958) (IV); vol.63 18053 (1965) (V); vol. 33 619 (VI).

ALBERT T. MEYERS, Primary Examiner F. E. WADDELL, Assistant ExaminerUNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,560,620 Dated February 2, 1971 Inventor-(s) Joseph Schol" t 8.1

It is certified that error appears in the above-identified pateni andthat said Letters Patent are hereby corrected as shown below:

In column 1, the upper right-hand corner of Formula A for "R" R line 55,for "athrom-" read a thromline 70, for "potency" read patenoy line 71,for "acitvator" read activa Column 2, line 31, for "lyti cactivity" readlytic activity lines 3H-35, for "effeotice" read effective line 61, for"activ or" read activity on line 62, for "icidence" read incidenceColumn 3, line 29 for "alJ ylene-bis-[3, +-dihydroiso-" read allcylenebis[3,H-dihydroiso- Column H, line 26 for "filitration" read filtrationline 28, for "diulent" read diluent Column 5, ll, for "product" readproduced line 25, for "methanolethyl" re methanol-ethyl line 31, for"procedude" read procedure lin for the title of Example 4, readl,l'-I'ETRAIIBTHY1ENEBIS[1,2,3,H-TEIR B-I'IL'IHOXYISOQUINOLINEJ.DIHYDROQiLDRIDE Column 6, line 52, after "p read (9.1) Column 7, line3, for "tetraisoquino-" read tetrahydroisoquinoline 13, after "suitable"read tablet 17, for "tetramethylene-" read tetrahydrolines 35, &7 and 60To make the solution isotonic under the heading "Ampoule"; line for "Perampoule" read Mg/Ampoule Column 9 line t, after "For A is" read a line12, after "A is" read a Signed and sealed this 7th day of September1971.

(SEAL) Attest:

EDNAHD M.FLETGI ER,JR.

Acting Commissioner of Pate

